Interfering RNA delivery system and uses thereof

Abstract

The invention provides a delivery system comprising a cell penetrating peptide, a polyarginine peptide, and an interfering RNA molecule. The system can be used for delivering interfering RNA molecules into a cell in vivo or in vitro. Therapeutic uses for the delivery system are also provided.

1. A method of treating an ocular disorder in a patient, comprising administering to the patient the interfering RNA delivery system comprising an interfering RNA molecule linked to a cell penetrating peptide (CPP)-Arg peptide, wherein the interfering RNA molecule can attenuate expression of a gene associated with the ocular disorder, and wherein the CPP-Arg peptide comprises the sequence as set forth in SEQ ID NO: 12. 
     
     
       2. The method of  claim 1 , wherein the ocular disorder is associated with ocular angiogenesis, dry eye, ocular inflammatory conditions, ocular hypertension, or glaucoma. 
     
     
       3. The method of  claim 1 , wherein the system is administered by intraocular injection, ocular topical application, subconjunctival injection, intravitreal injection, anterior or posterior juxtascleral injection, intravenous injection, oral administration, intramuscular injection, intraperitoneal injection, transdermal application, intranasal application, or transmucosal application. 
     
     
       4. A method of delivering an interfering RNA molecule into a cell, comprising:
 (a) attaching an interfering RNA molecule to a CPP-Arg peptide, thereby forming an interfering RNA delivery system, wherein the CPP-Arg peptide comprises the sequence as set forth in SEQ ID NO: 12; and 
 (b) administering the interfering RNA delivery system to the cell under conditions suitable for the system to enter the cell. 
 
     
     
       5. The method of  claim 4 , wherein release of the system from the endosome inside the cell is enhanced in the presence of an HA2 peptide. 
     
     
       6. The method of  claim 5 , wherein the HA2 peptide is introduced into the cell by administration of a CPP-HA2 peptide. 
     
     
       7. The method of  claim 4 , wherein the CPP-Arg peptide further comprises the HA2 peptide. 
     
     
       8. The method of  claim 4 , wherein the interfering RNA molecule is linked to the CPP-Arg peptide via a covalent bond. 
     
     
       9. The method of  claim 4 , wherein the interfering RNA molecule is a siRNA, miRNA, or shRNA. 
     
     
       10. The method of  claim 4 , wherein the CPP has an amino acid sequence in the forward or reverse direction. 
     
     
       11. The method of  claim 10 , wherein the CPP comprises at least one D-amino acid.